Difference between revisions of "Stains"

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This article deals with '''stains'''. H&E isn't the only stain out there...
[[Image:Corpora amylacea high mag.jpg|thumb|300px|[[Hematoxylin and eosin stain]] of benign [[prostate gland]].]]
This article deals with '''stains'''. The H&E stain is only one of many stains that are used in pathology.


==Where to start...==
Non-H&E stains are often referred to as '''''special stains'''''.
===Principles===
 
=Where to start...=
==Principles==
When considering additional (i.e. special) stains one should (in order) do the following:<ref>LAE. 13 July 2010.</ref>
When considering additional (i.e. special) stains one should (in order) do the following:<ref>LAE. 13 July 2010.</ref>
# Make sure one has exhausted the clinical history; history is considered the best special stain.
# Make sure one has exhausted the clinical history; history is considered the best special stain.
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# Molecular testing, electron microscopy.
# Molecular testing, electron microscopy.


===Common stains===
==Common stains==
# H&E stain.
# [[H&E stain]].
# PAS stain.
# [[PAS stain]].
# PAS-D stain.
# [[PAS-D stain]].
# AFB stain.
# [[AFB stains]], e.g. [[Ziehl-Neelsen stain]].
# Congo red.
# [[Congo red]].
# GMS stain.
# [[GMS stain]].
# Gram stain.
# [[Gram stain]].


==Immunohistochemistry==
=Immunohistochemistry=
{{main|Immunohistochemistry}}
{{main|Immunohistochemistry}}
===General===
==General==
*Abbreviated ''IHC''.
*Abbreviated ''IHC''.


===Interpretation===
==Interpretation==
Simple version:
Simple version:
*Positive is (usually): ''brown''.
*Positive is (usually): ''brown''.
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*The edge of the tissue may have light staining - ''edge effect''.
*The edge of the tissue may have light staining - ''edge effect''.
*If everything is brown... suspect that it didn't work.
*If everything is brown... suspect that it didn't work.
*In some situations you're blessed with an ''internal control'', e.g. in renal tumours CD10 will stain [[RCC]] and the ''proximal tubule'', in GISTs - CD117 the mast cells are positive.
*In some situations you're blessed with an ''internal control'', e.g. in renal tumours CD10 will stain [[RCC]] and the ''proximal tubule'', in GISTs - CD117 the [[mast cell]]s are positive.


==Work-up of infection==
=Work-up of infection=
It often not possible to be definitive by staining.<ref>{{cite journal |author=Woods GL, Walker DH |title=Detection of infection or infectious agents by use of cytologic and histologic stains |journal=Clin. Microbiol. Rev. |volume=9 |issue=3 |pages=382-404 |year=1996 |month=July |pmid=8809467 |pmc=172900 |doi= |url=http://cmr.asm.org/cgi/pmidlookup?view=long&pmid=8809467}}</ref>
It often not possible to be definitive by staining.<ref>{{cite journal |author=Woods GL, Walker DH |title=Detection of infection or infectious agents by use of cytologic and histologic stains |journal=Clin. Microbiol. Rev. |volume=9 |issue=3 |pages=382-404 |year=1996 |month=July |pmid=8809467 |pmc=172900 |doi= |url=http://cmr.asm.org/cgi/pmidlookup?view=long&pmid=8809467}}</ref>


Basic panel:
Basic panel:
*Gram stain - for bacteria.
*Gram stain - for bacteria.
*GMS stain - fungal stain.
*GMS stain - [[fungi|fungal]] stain.
*PAS ''or'' PAS-D - for fungi (???).
*PAS (''or'' PAS-D) - fungal stain.


===Fungi===
==Fungi==
{{main|Microorganisms}}
{{main|Fungi}}


==Specific stains==
Fungi are a type of [[microorganisms]].  They are seen by pathologist every once in a while.
 
=Specific stains=
What follows is a big list... of stains.
What follows is a big list... of stains.


==Haematoxylin & Eosin stain==
==Haematoxylin and eosin stain==
===General===
*Abbreviated ''H&E''.
*Abbreviated ''H&E''.
*Standard bearer in most pathology departments.
{{Main|Hematoxylin and eosin stain}}
 
===Intepretation===
*Blue (haematoxylin) = nucleus.
*Pink (eosin) = cytoplasm.


==Haematoxylin phyloxin saffron stain==
==Haematoxylin phyloxin saffron stain==
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===Interpretation===
===Interpretation===
*Haematoxylin = blue -- stains nucleus.
*Haematoxylin = blue -- stains nucleus.
*Phyloxin = pink.
*Phyloxin = pink -- stains muscle and cytoplasm.
*Saffron = yellow -- stains collagen.
*Saffron = yellow -- stains collagen.
*An alternative to H&E stain.
*An alternative to H&E stain.
**Fibrosis is easier to see on HPS than H&E... as one can see the collagen.
**Fibrosis is easier to see on HPS than H&E... as one can see the collagen.


====Images====
<gallery>
Image:Perineural_invasion_prostate_high_mag.jpg | Perineural invasion - prostate - HPS stain (WC)
Image:Meningioma_high_mag.jpg | Meningioma - HPS stain (WC)
Image:Endoneurial_fibrosis_-_very_high_mag_-_cropped.jpg | Endoneurial fibrosis - HPS stain (WC)
</gallery>
==Periodic acid Schiff stain==
==Periodic acid Schiff stain==
*Abbreviated ''PAS''.
*Abbreviated ''PAS''.
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*Kidney biopsies, medical.
*Kidney biopsies, medical.
*Liver biopsies, medical.
*Liver biopsies, medical.
**Positive in ''alpha-1 antitrypsin deficiency''.


===Utility===
===Utility===
*Stains - basement membranes, fungi, glycogen, mucin  
*Stains - lipofuscin,<ref name=pmid5463681 >{{cite journal |author=Kovi J, Leifer C |title=Lipofuscin pigment accumulation in spontaneous mammary carcinoma of A/Jax mouse |journal=J Natl Med Assoc |volume=62 |issue=4 |pages=287–90 |year=1970 |month=July |pmid=5463681 |pmc=2611776 |doi= |url=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2611776/pdf/jnma00512-0077.pdf}}</ref> basement membranes, fungi, glycogen, (neutral) mucin.


===Interpretation===
===Interpretation===
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*Blue = nuclei.
*Blue = nuclei.


Notes: <ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF]</ref>
Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF]</ref>
====Image====
<gallery>
Image:Metanephric_adenoma_high_mag.jpg | Metanephric adenoma - PAS (WC)
File:Liver biopsy of glycogen storage disorder PAS positive.jpg | Liver biopsy with glycogen storage disease - PAS (WC)
File:Esophageal candidiasis (2) PAS stain.jpg | Esophageal candidiasis - PAS (WC)
</gallery>
 
==Periodic acid Schiff fungal stain==
*Abbreviated ''PASF''.
 
===Primary application===
*Look for [[fungi|fungal organisms]].
 
===Interpretation===
*Light purple = fungi.
*Light green = background.
*Washed-out light purple = Gram positive bacilli.
 
Note:
*This is much improved over the ''PAS'' in the context of skin, as the background is similar to the fungal organisms.
 
<gallery>
Image:Fungal_infection_-_PASF_-_very_high_mag.jpg | Fungal organisms. PASF stain.
</gallery>
 
==Periodic acid Schiff with diastase==
==Periodic acid Schiff with diastase==
*Abbreviated: ''PAS-D'' and ''PASD''.
*Abbreviated: ''PAS-D'' and ''PASD''.
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===General===
===General===
*Diastase digests the glycogen.
*Diastase digests the glycogen.
*"PAS diastase resistant"<ref name=pmid6189389>{{Cite journal  | last1 = Qizilbash | first1 = A. | last2 = Young-Pong | first2 = O. | title = Alpha 1 antitrypsin liver disease differential diagnosis of PAS-positive, diastase-resistant globules in liver cells. | journal = Am J Clin Pathol | volume = 79 | issue = 6 | pages = 697-702 | month = Jun | year = 1983 | doi =  | PMID = 6189389 }}</ref> ''implies'' PASD +ve and [[PAS stain|PAS]] +ve.


===Use===
===Use===
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Notes: <Ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF]</ref>
Notes: <Ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF]</ref>
===Interpretation===
*Light purple = fungi.
*Light blue/pink = background. ???


==Gomori methenamine-silver stain==
==Gomori methenamine-silver stain==
*Abbrev. ''GMS''.
*Abbreviated ''GMS''.
 
Note:
*''GMS'' is "Grocott's methenamine Silver" according to WMSP.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>
 
===Use===
*Useful for fungi.  
*Useful for fungi.  
**Pneumocystis jirovecii - cause of ''PCP'' - pneumocystic pneumonia.
**Pneumocystis jirovecii - cause of [[pneumocystis pneumonia]] (PCP).
**Histoplasma - cause of histoplasmosis.
**Histoplasma - cause of [[histoplasmosis]].
***Histoplasma = black, round balls.
***Histoplasma = black, round balls.


Notes: ''GMS'' is "Grocott's methenamine Silver" according to WMSP.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>
====Image====
 
<gallery>
Image:Histoplasma_in_granuloma_gms.jpg | GMS showing histoplasma (WC/Nephron)
</gallery>
==Acid-fast bacilli stains==
==Acid-fast bacilli stains==
*Abbreviated: ''AFB''.
*Abbreviated: ''AFB''.


There are several AFB stains:
There are several AFB stains:
*Ziehl-Neelson stain - used to look for Mycobacterium tuberculosis.
*Ziehl-Neelson stain - used to look for ''[[Mycobacterium tuberculosis]]''.
*Fite stain - used to look for Mycobacterium laprae.<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF]</ref>
*Fite stain - used to look for ''[[Mycobacterium leprae]]''<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF]. Accessed on: 19 May 2011.</ref> and preferred stain for ''[[Mycobacterium avium complex]]''.<ref name=pmid21327589>{{cite journal |authors=Ilyas S, Youssef D, Chaudhary H, Al-Abbadi MA |title=Myocbacterium-avium intracellulare associated inflammatory pseudotumor of the anterior nasal cavity |journal=Head Neck Pathol |volume=5 |issue=3 |pages=296–301 |date=September 2011 |pmid=21327589 |pmc=3173547 |doi=10.1007/s12105-011-0248-0 |url=}}</ref>
*Auramine rhodamine.
*Auramine-rhodamine stain.


===Ziehl-Neelson stain===
===Ziehl-Neelsen stain===
*Most popular acid-fast bacilli stain.
*Most popular acid-fast bacilli stain.
*Stains other mycobacteria -- not specific for tuberculosis.
*Stains other mycobacteria -- not specific for tuberculosis.
**Stains nocardia.<ref>[http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html]</ref>
**Stains ''Nocardia''.<ref>URL: [http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html]. Accessed on: 19 May 2011.</ref>


====Image====
<gallery>
Image:Mycobacterium_tuberculosis_Ziehl-Neelsen_stain_02.jpg | ZN stain. (WC/CDC)
</gallery>
===Fite stain===
===Fite stain===
Interpretation:
Interpretation:
*Red = AFB.
*Red = AFB.
*Blue = background.
*Blue = background.
===Auramine-rhodamine stain===
*Fluorescent stain.
====Image====
<gallery>
Image:Cryptosporidium_parvum_auramine-rhodamine_labeled.jpg | AR stain. (WC/CDC)
</gallery>
===Kinyoun stain===
*Another AFB stain<ref name=pmid7536216>{{Cite journal  | last1 = Kehl | first1 = KS. | last2 = Cicirello | first2 = H. | last3 = Havens | first3 = PL. | title = Comparison of four different methods for detection of Cryptosporidium species. | journal = J Clin Microbiol | volume = 33 | issue = 2 | pages = 416-8 | month = Feb | year = 1995 | doi =  | PMID = 7536216 }}</ref> - useful for [[cryptosporidiosis]] and [[microsporidiosis]].<ref name=pmid9003613>{{Cite journal  | last1 = Ignatius | first1 = R. | last2 = Lehmann | first2 = M. | last3 = Miksits | first3 = K. | last4 = Regnath | first4 = T. | last5 = Arvand | first5 = M. | last6 = Engelmann | first6 = E. | last7 = Futh | first7 = U. | last8 = Hahn | first8 = H. | last9 = Wagner | first9 = J. | title = A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens. | journal = J Clin Microbiol | volume = 35 | issue = 2 | pages = 446-9 | month = Feb | year = 1997 | doi =  | PMID = 9003613 }}
</ref>


==Congo red stain==
==Congo red stain==
===Use===
*Used to look for [[amyloid]].
*Used to look for [[amyloid]].
*Mnemonic: ''CRAP'' = congo red amyloid protein.
**Mnemonic: ''CRAP'' = congo red amyloid protein.
*An alternate stain for amyloid is ''Thioflavin T''.
*An alternate stain for amyloid is ''Thioflavin T''.


Interpretation:
Note:
*[[Cutting|Thick sections]] (~10 micrometers) are considered a requirement for the stain to work properly.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/congo_red_bennhold.htm http://www.ihcworld.com/_protocols/special_stains/congo_red_bennhold.htm]. Accessed on: 26 January 2012.</ref>
**If the section is too thin... it doesn't work.
 
===Interpretation===
*Amyloid = pink/red.
*Amyloid = pink/red.
*Nuclei = blue.
*Nuclei = blue.


Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF]</ref>
Ref.:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF]. Accessed on: 4 December 2010.</ref>
 
====Image====
<gallery>
Image:Cerebral_amyloid_angiopathy_-_very_high_mag.jpg | Congo red staining in [[cerebral amyloid angiopathy]]. (WC)
</gallery>
 
==Thioflavin T stain==
===Use===
*Used to look for [[amyloid]].
 
===Interpretation===
*Amyloid = green.
 
Image: [http://inano.au.dk/research/annual-reports/annual-report-2004/7-cases/protein-fibrils/ Amyloid (inano.au.dk)].
 
==Gram stain==
==Gram stain==
Use:
===Use===
*"It is useless for finding bacteria."<ref>SMH handout.</ref>
*"It is useless for finding bacteria."<ref>St. Michael's Hospital - Stains Handout.</ref>
**I suppose this is as you find 'em on H&E... or microbiology finds 'em.
**If they are to be seen... they'll be visible on H&E.
 
Note:
*Microbiology is better at finding organisms than pathology.
**They have one significant advantage -- if a small amount of bugs are present... they grows into a large (obviously visible) colony.
 
====DDx for common patterns====
A short list of bacteria and their characteristics:<ref>URL: [http://www.atsu.edu/faculty/chamberlain/Website/pnebact.htm http://www.atsu.edu/faculty/chamberlain/Website/pnebact.htm]. Accessed on: 7 May 2013.</ref>
{| class="wikitable sortable"
! Shape\Gram stain
! Positive
! Negative
! Variable or negative
|-
| Bacilli
| Clostridium difficile, Bacillus anthracis, Nocardia spp.
| Escherichia coli, [[Helicobacter pylori]], Yersinia pestis, Hemophilus influenzae
| [[Tuberculosis|Mycobacterium tuberulosis]], Legionella pneumophila<ref>URL: [http://meded.ucsd.edu/isp/1999/CAP/legion.html http://meded.ucsd.edu/isp/1999/CAP/legion.html]. Accessed on: 7 May 2013.</ref>
|-
| Cocci
| Streptococcus pneumoniae, Staphylococcus aureus
| Neisseria meningitidis, Moraxella catarrhalis
|
|}


===Interpretation===
===Interpretation===
*Blue = Gram positive organisms.
*Purple (or blue) = Gram positive organisms.
*Red = Gram negative organisms, nuclei
*Red = Gram negative organisms, nuclei.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF]. Accessed on: 7 May 2013.</ref>
*Yellow = background.
*Yellow = background.


Notes:
Notes:
*Many of the bacteria are quite small relative to lymphocytes; ''Escherichia coli'' is 1-2 micrometers long x 0.25 micrometers in diameter.<ref>[http://www.lpi.usra.edu/publications/slidesets/marslife/slide_27.html http://www.lpi.usra.edu/publications/slidesets/marslife/slide_27.html]</ref>
*Many of the bacteria are quite small relative to lymphocytes; ''Escherichia coli'' is 1-2 micrometers long x 0.25 micrometers in diameter.<ref>URL: [http://www.lpi.usra.edu/publications/slidesets/marslife/slide_27.html http://www.lpi.usra.edu/publications/slidesets/marslife/slide_27.html].</ref>
*Epithelial cell nuclei & stromal cell nuclei may stain red.
*Epithelial cell nuclei & stromal cell nuclei may stain red.
*Memory device: '''p'''urple = '''p'''ositive.


Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF]</ref>
====Images====
 
<gallery>
Image:Gram_stain_01.jpg | Gram positive cocci. (WC)
Image:Gram_Stain_Anthrax.jpg | Gram positive rods - anthrax. (WC)
</gallery>
==Luxol fast blue stain==
==Luxol fast blue stain==
*Abbreviated ''LFB''.
*Abbreviated ''LFB''.
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===Intepretation===
===Intepretation===
*Blue = myelinated fibers (contain lipoproteins).
*Blue = myelinated fibers (contain lipoproteins), lipofuscin.<ref>MUN. 26 November 2010.</ref>
**Lack of blue suggests demyelination.
**Lack of blue (where it ought to be) = demyelination.
*Purple = nerve cell (e.g. neuron).
*Purple = nerve cell (e.g. neuron).
*Neutrophils = pink.
*[[Neutrophil]]s = pink.


Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF]</ref>
Ref.:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF]</ref>
 
====Image====
<gallery>
Image:Globus_pallidus_and_putamen_-_very_low_mag.jpg | Globus pallidus and putamen - H&E-LFB. (WC)
File:LFB_CNS_cortex_supratentorial.jpg | Normal cortex - LFB only. (WC/jensflorian)
File:LFB_CNS_cortex_grey-white_matter_junction.jpg | White-grey matter junction - LFB. (WC/jensflorian)
</gallery>


==Giemsa stain==
==Giemsa stain==
===Use===
===Use===
*Useful for finding mast cells.
*Useful for finding [[mast cell]]s.
*Useful for finding donovan bodies and leishmania.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: April 6, 2009.</ref>
*Useful for finding ''[[Donovan bodies]]'' and ''[[Leishmania]]''.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: April 6, 2009.</ref>


===Interpretation===
===Interpretation===
*Tissue is light blue/green.
*Tissue is light blue/green.
*Goblet cells are purple.<ref>URL: [http://www.kennedy.ox.ac.uk/facilities/histology/histology-information http://www.kennedy.ox.ac.uk/facilities/histology/histology-information]. Accessed on: 17 August 2015.</ref>
Image:
*[http://path.upmc.edu/cases/case196/images/figure12.jpg Giemsa - colon (amser.org)].<ref>URL: [http://amser.org/index.php?P=AMSER--ResourceFrame&resourceId=6018 http://amser.org/index.php?P=AMSER--ResourceFrame&resourceId=6018]. Accessed on: 17 August 2015.</ref>


==Reticulin stain==
==Reticulin stain==
===Use===
===Use===
*Liver biopsy, medical.
*Liver biopsy, medical.
**Demonstrates the reticular fibers (in cirrhosis the fibers are disrupted).
**Demonstrates the reticular fibers (in [[cirrhosis]] the fibers are disrupted).
*Before IHC, reticulin was used to differentiate ''sarcomas'' from ''carcinomas'':<ref name=pmid13536209>{{cite journal |author=MACKENZIE DH |title=Reticulin patterns in the diagnosis of carcinomas and sarcomas |journal=Br. J. Cancer |volume=12 |issue=1 |pages=14–9 |year=1958 |month=March |pmid=13536209 |pmc=2074006 |doi= |url=}}</ref>
**Sarcomas have reticulin around ''each'' cell.
**Carcinomas have reticulin around clusters of cells.
*Commonly used in neuropathology.
** In adenoma, reticulin highlights the lost acinar structure of normal pituitary gland.
** Paraganglioma (Zellballen architecture)
** Separating schwannoma (basement membrane around each cell) from meingioma in cerebellopontine angle.
** Separating desmoplastic medulloblastoma from classic/anaplastic forms.
 


===Interpretation===
===Interpretation===
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Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF]</ref>
Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF]</ref>


==Cresyl violet==
====Images====
<gallery>
Image:Liver_reticulin.jpg | [[Liver]]. Reticulin stain. (WC)
Image:Hepatic_adenoma_high_mag_reticulin.jpg | [[Hepatic adenoma]]. Reticulin stain. (WC)
File:Zellballen paraganglioma.jpg | Reticulin stain highlighting the "Zellballen" architecture of paraganglioma. (WC/jensflorian)
File:Desmoplastic medulloblastoma reticulin stain pale island.jpg | Reticulin staina round the "pale islands" of a desmoplastic medulloblastoma. (WC/jensflorian)
</gallery>
 
==Cresyl violet stain==
===Use===
*Used at some places (e.g. SMH) to look for Helicobacter organisms.
 
===Interpretation===
*Everything is shades of blue.
*Everything is shades of blue.
*Used at some places (e.g. SMH) to look for Helicobacter organisms.
**Helicobacter stains blue.
**Helicobacter stains blue.


==Prussian blue stain==
==Prussian blue stain==
*AKA ''Perl's iron stain''.
*AKA ''Perl's iron stain''.
===Use===
*Useful for iron and hemosiderin; useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
*Useful for iron and hemosiderin; useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
===Interpretation===
*Blue = iron.


Image:
Image:
Line 199: Line 340:
*Described well by ''vetmed.vt.edu''.<ref>Prussian blue stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm [http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm]. Accessed on: 5 May 2010.</ref>
*Described well by ''vetmed.vt.edu''.<ref>Prussian blue stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm [http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm]. Accessed on: 5 May 2010.</ref>
*DDx of brown pigment: Fontana-Masson (melanin), Kluver-Barrera stain (lipofuscin).
*DDx of brown pigment: Fontana-Masson (melanin), Kluver-Barrera stain (lipofuscin).
====Images====
<gallery>
Image:Hemosiderosis_high_mag.jpg | Liver [[hemosiderosis]]. Prussian blue stain. (WC/Nephron)
File:Siderophage iron stain CSF.jpg | CSF Siderophages in subarachnoid hemorrhage. (WC/jensflorian)
</gallery>


==Kluver-Barrera stain==
==Kluver-Barrera stain==
Line 209: Line 356:
*Useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
*Useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
**Stains lipofuscin.
**Stains lipofuscin.
*Useful to detect demyelinating lesions in the CNS.
Notes:
*[[PAS stain|PAS]] also stains lipofuscin and is more commonly available.


===Interpretation===
===Interpretation===
Line 216: Line 367:
*Described well by ''vetmed.vt.edu''.<ref>Kluver-Barrera stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm]. Accessed on: 5 May 2010.</ref>
*Described well by ''vetmed.vt.edu''.<ref>Kluver-Barrera stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm]. Accessed on: 5 May 2010.</ref>
*DDx of brown pigment: Fontana-Masson (melanin), Prussian blue stain (hemosiderin).
*DDx of brown pigment: Fontana-Masson (melanin), Prussian blue stain (hemosiderin).
<gallery>
File:MS Demyelinisation KB 10x.jpg | Encephalomyelitis disseminata (Klüver-Barrera)
</gallery>


==Oil red 0 stain==
==Oil red O stain==
===Use===
===Use===
*Stain adipose tissue.
*Stains adipose tissue.
*Corroborate diagnosis of [[lipoid pneumonia]].<ref name=pmid25374742>{{Cite journal  | last1 = Yampara Guarachi | first1 = GI. | last2 = Barbosa Moreira | first2 = V. | last3 = Santos Ferreira | first3 = A. | last4 = Sias | first4 = SM. | last5 = Rodrigues | first5 = CC. | last6 = Teixeira | first6 = GH. | title = Lipoid pneumonia in a gas station attendant. | journal = Case Rep Pulmonol | volume = 2014 | issue =  | pages = 358761 | month =  | year = 2014 | doi = 10.1155/2014/358761 | PMID = 25374742 }}</ref>
*Screen for [[GERD]] - positive staining seen in macrophages from [[BAL]] specimens.<ref name=pmid20466562>{{Cite journal  | last1 = Hopkins | first1 = PM. | last2 = Kermeen | first2 = F. | last3 = Duhig | first3 = E. | last4 = Fletcher | first4 = L. | last5 = Gradwell | first5 = J. | last6 = Whitfield | first6 = L. | last7 = Godinez | first7 = C. | last8 = Musk | first8 = M. | last9 = Chambers | first9 = D. | title = Oil red O stain of alveolar macrophages is an effective screening test for gastroesophageal reflux disease in lung transplant recipients. | journal = J Heart Lung Transplant | volume = 29 | issue = 8 | pages = 859-64 | month = Aug | year = 2010 | doi = 10.1016/j.healun.2010.03.015 | PMID = 20466562 }}</ref>
*Uncommon.


Notes:
Notes:
*Must be done on fresh tissue, i.e. it cannot be fixed in formulin.
*Must be done on fresh tissue, i.e. it cannot be fixed in [[formalin]].
 
===Interpretation===
*Red = fat.
 
====Images====
<gallery>
Image:Differentiated_3T3-L1_Cell_line_stained_with_Oil_O_Red.jpg | Oil red O stain. (WC)
</gallery>


==Warthin-Starry stain==
==Warthin-Starry stain==
Background
Background:
*Developed by a bunch of pathologists in Michigan to look for spirochetes<ref>[http://www.merriam-webster.com/medical/warthin]</ref>
*Developed by a bunch of pathologists in Michigan to look for spirochetes.<ref>URL: [http://www.merriam-webster.com/medical/warthin http://www.merriam-webster.com/medical/warthin]. Accessed on: 17 August 2010.</ref>


===Use===
===Use===
*Find spirochetes, e.g. syphilis<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: April 6, 2009.</ref>
*Find spirochetes, e.g. [[syphilis]] (Treponema pallidum),<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: April 6, 2009.</ref> cat-scratch disease (Bartonella henselae).
*Find Helicobacter spp., e.g. Helicobacter pylori -- Mount Sinai Hospital<ref>[http://www.dako.co.uk/index/prod_search/prod_products.htm?productareaid=41&baseprodidver=A224462007 http://www.dako.co.uk/index/prod_search/prod_products.htm?productareaid=41&baseprodidver=A224462007]</ref>
*Find Helicobacter spp., e.g. Helicobacter pylori -- Mount Sinai Hospital.<ref>[http://www.dako.co.uk/index/prod_search/prod_products.htm?productareaid=41&baseprodidver=A224462007 http://www.dako.co.uk/index/prod_search/prod_products.htm?productareaid=41&baseprodidver=A224462007]</ref>


Interpretation:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAIN029.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAIN029.html]</ref>
Interpretation:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAIN029.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAIN029.html]</ref>
Line 236: Line 401:
*Background - yellow.
*Background - yellow.


Image:  
====Image====
*http://commons.wikimedia.org/wiki/File:Pylorigastritis.jpg Helicobacter gastritis - Warthin-Starry stain (WC)].
<gallery>
Image:Pylorigastritis.jpg | Helicobacter gastritis - Warthin-Starry stain. (WC)
</gallery>
Notes:
*Considered a "dirty" stain - picks-up junk in the background.<ref>DB. 4 August 2010.</ref>
 
==Dieterle stain==
Considered a variant of the ''Steiner stain''.<ref>URL: [http://www.mayomedicallaboratories.com/test-catalog/Overview/80327 http://www.mayomedicallaboratories.com/test-catalog/Overview/80327]. Accessed on: 8 August 2010.</ref>
===Use===
*Find spirochetes, e.g. [[syphilis]] (Treponema pallidum),<ref name=Ref_WMSP455>{{Ref WMSP|455}}</ref> donovan bodies (leishmaniasis),<ref>URL: [http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain]. Accessed on: 4 August 2010.</ref> Helicobacter pylori and Bartonella henselae (Cat-scratch disease).<ref>URL: [http://www.mayomedicallaboratories.com/test-catalog/Overview/80327 http://www.mayomedicallaboratories.com/test-catalog/Overview/80327]. Accessed on: 8 August 2010.</ref>
 
===Interpretation===
*Spirochetes - black.
*Background - yellow.
 
====Images====
<gallery>
Image:Treponema_pallidum_-_very_high_mag_-_extreme_crop.jpg | Dieterle stain - T. pallidum. (WC)
</gallery>
www:
*[http://pathmicro.med.sc.edu/trepo.jpg Treponema (med.sc.edu)].
*[http://pathmicro.med.sc.edu/fox/spiro-neisseria.htm Spirochetes - several images (med.sc.edu)].


==Bielschowsky stain==
==Bielschowsky stain==
Line 251: Line 437:
*Yellow/brown = other.
*Yellow/brown = other.


Notes: <ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF]</ref>
Ref.: <ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF]</ref>
==Mucicarmine==
====Image====
<gallery>
Image:Cerebellum_-_biel_-_very_high_mag.jpg | Bielschowsky stain. (WC/Nephron)
</gallery>
==Mucicarmine stain==
*Stains some mucins... uses the dye ''carmine''.
*Stains some mucins... uses the dye ''carmine''.


===Use===
===Use===
*Identify mucin.
*Identify mucin.
*Malignant cells that produce mucin... carcinomas.<ref>APBR P.681 (Q25).</ref>
*Malignant cells that produce mucin... carcinomas.<ref name=Ref_APBR681>{{Ref APBR|681 (Q25)}}</ref>


===Interpretation===  
===Interpretation===  
Line 265: Line 455:
**Red = mucin.<ref>[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg]</ref>
**Red = mucin.<ref>[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg]</ref>


Image:
====Images====
*[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/Pages/page6.htm Mucicarmine stained bowel (medschool.lsuhsc.edu)]. [http://www.nature.com/modpathol/journal/v14/n5/fig_tab/3880332f4.html Mucicarmine stained pancreatic adenosquamous carcinoma (nature.com)].
<gallery>
Image:Cryptococcosis of lung in patient with AIDS. Mucicarmine stain 962 lores.jpg | [[Cryptococcosis]]. Mucicarmine stain. (WC/CDC)
</gallery>
www:
*[http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/Pages/page6.htm Mucicarmine stained bowel (medschool.lsuhsc.edu)].  
*[http://www.nature.com/modpathol/journal/v14/n5/fig_tab/3880332f4.html Mucicarmine stained pancreatic adenosquamous carcinoma (nature.com)].


==Alcian blue==
==Alcian blue stain==
===General===
===General===
*Stains acidic mucin (pH=2.5); '''A'''lcian blue = '''A'''cidic.
*Stains acidic mucin (pH=2.5); '''A'''lcian blue = '''A'''cidic.
**A variant uses pH=1.0.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>
**A variant uses pH=1.0.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>


Note:
*''Alcian blue'' (not otherwise specified) usu. refers to the pH=2.5.<ref>URL: [http://www.pathologyoutlines.com/topic/stainsalcianblue.html http://www.pathologyoutlines.com/topic/stainsalcianblue.html]. Accessed on: 11 October 2012.</ref>
===Use===
*Identify ''[[intestinal metaplasia]]'' in the [[intestinal metaplasia of the stomach|stomach]] and [[Barrett esophagus|esophagus]] -- goblets = blue.
Note:
*Esophageal submucosal glands - alcian blue positive.
===Interpretation===
*Blue = acidic mucins.<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF]. Accessed on: 20 December 2011.</ref>
Notes:
*Mucin stains:
**[[Alcian blue stain]], [[PASD stain]], [[Mucicarmine stain]].
====Image====
<gallery>
Image:Barrett's_mucosa,_PAS-Alcian_blue_stain.jpg | [[Barrett's esophagus|Barrett's type mucosa]]. Alcian blue stain. (WC/AFIP)
Image:Barrett's_mucosa,_higher_magnification,_Alcian_blue_stain_.jpg | [[Barrett's esophagus|Barrett's type mucosa]]. Alcian blue stain. (WC/AFIP)
</gallery>
==Sodium sulphate-alcian blue stain==
:''Sulfated alcian blue'' (abbreviated '''SAB'') redirects here.
===Use===
===Use===
*Identify ''intestinal metaplasia'' in the [[stomach]] -- goblets = blue.
*Identify [[amyloid]].<ref name=pmid55419>{{Cite journal  | last1 = Pomerance | first1 = A. | last2 = Slavin | first2 = G. | last3 = McWatt | first3 = J. | title = Experience with the sodium sulphate-Alcian Blue stain for amyloid in cardiac pathology. | journal = J Clin Pathol | volume = 29 | issue = 1 | pages = 22-6 | month = Jan | year = 1976 | doi =  | PMID = 55419 }}</ref><ref>URL: [http://www.polyrnd.com/products/reagent-assembly-kits/conventional/amyloid-stain---sulfated-alcian-blue-(sab).aspx http://www.polyrnd.com/products/reagent-assembly-kits/conventional/amyloid-stain---sulfated-alcian-blue-(sab).aspx]. Accessed on: October 15, 2014.</ref>
*Less [[specificity|specific]] than [[congo red stain|congo red]] but equally [[sensitivity|sensitive]].
*Does not require polarized light.


===Interpretation===
===Interpretation===
*Blue = acidic mucins.
*Green = amyloid.<ref name=pmid55419/>
**Other things that are green: [[mast cell]]s, mucoid degeneration, basophilic myofibre degeneration, califications.
*Yellow = background.
 
Image:
*[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC475937/figure/F1/ SAB (nih.gov)].


Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF]</ref>
==Movat's stain==
==Movat's stain==
===Use===
===Use===
Line 296: Line 521:


How to remember? A.: Primary colours (red, blue, yellow) + black.
How to remember? A.: Primary colours (red, blue, yellow) + black.
==Masson's trichrome==
 
===General use===
====Images====
<gallery>
Image:Cardiac_amyloidosis_very_high_mag_movat.jpg | [[Cardiac amyloidosis]] - Movat stain. (WC/Nephron)
Image:Cystic_medial_degeneration_-_movat_-_low_mag.jpg | [[Cystic medial degeneration]] - Movat stain - low mag. (WC/Nephron)
Image:Cystic_medial_degeneration_-_movat_-_intermed_mag.jpg | Cystic medial degeneration - Movat stain - intermed. mag. (WC/Nephron)
Image:Cystic medial degeneration - movat - high mag.jpg | Cystic medial degeneration - Movat stain - high mag. (WC/Nephron)
</gallery>
 
==Masson's trichrome stain==
*Should '''not''' be confused with the ''[[Mallory trichrome stain]]''.
*May be referred to as ''[[trichrome stain]]''.
===General===
*Collagen vs. muscle.
*Collagen vs. muscle.
===Site===
*Kidney Bx (to assess for fibrosis).
*Liver Bx (to assess for [[cirrhosis]]).


===Interpretation===
===Interpretation===
Line 308: Line 540:
*Red = muscle (smooth muscle actin).
*Red = muscle (smooth muscle actin).
*Baby blue = collagen.
*Baby blue = collagen.
*Blue gray = [[amyloid]].<ref name=pmid33002919>{{cite journal |authors=Kunnath-Velayudhan S, Larsen BT, Coley SM, De Michele S, Santoriello D, Colby TV, Bhagat G, Saqi A |title=Masson Trichrome and Sulfated Alcian Blue Stains Distinguish Light Chain Deposition Disease From Amyloidosis in the Lung |journal=Am J Surg Pathol |volume=45 |issue=3 |pages=405–413 |date=March 2021 |pmid=33002919 |doi=10.1097/PAS.0000000000001593 |url=}}</ref>


Notes: <Ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF]</ref>
Notes: <Ref>ULR: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF]. Accessed on: 2 November 2011.</ref>


===Elastic trichrome===
===Elastic trichrome stain===
General:
General:
*"Elastic trichrome" is one important variant of ''Masson's trichrome''.
*"Elastic trichrome" is one important variant of ''Masson's trichrome''.
Line 318: Line 551:
*Black = nuclei and '''elastin'''.
*Black = nuclei and '''elastin'''.


==HOPS stain==
==Mallory trichome stain==
Name is an acronym for... ''haematoxylin'', ''orcein'', ''phyloxin'' and ''saffron''.<ref>{{cite journal |author=Perry JR, Bilbao JM, Gray T |title=Fatal basilar vasculopathy complicating bacterial meningitis |journal=Stroke |volume=23 |issue=8 |pages=1175–8 |year=1992 |pmid=1636194 |doi=}} [http://stroke.ahajournals.org/cgi/reprint/23/8/1175.pdf Free Full Text].</ref>
*Should '''not''' be confused with ''[[Masson trichrome stain]]''.
*May be referred to as ''[[trichrome stain]]''.
 
===General===
*Collagen vs. muscle.
*May be done with elastin.
 
===Site===
*Kidney Bx (to assess for fibrosis).
**Considered better than the [[Masson trichrome stain]].
*Liver Bx (to assess for [[cirrhosis]]).
*Cardiovascular/lung (to see differentiate the layers of the arteries, and arteries from veins).
 
===Interpretation===
*Black = nuclei.
*Red = muscle (smooth muscle actin).
*Green = collagen.
 
====Image====
<gallery>
Image:Cirrhosis_high_mag.jpg | [[Cirrhosis]]. Mallory trichrome. (WC/Nephron)
</gallery>
==Haematoxylin orcein phyloxin saffron stain==
*Abbreviated ''HOPS''.<ref name=pmid1636194>{{cite journal |author=Perry JR, Bilbao JM, Gray T |title=Fatal basilar vasculopathy complicating bacterial meningitis |journal=Stroke |volume=23 |issue=8 |pages=1175–8 |year=1992 |pmid=1636194 |doi=}} [http://stroke.ahajournals.org/cgi/reprint/23/8/1175.pdf Free Full Text].</ref>
*It should ''not'' be confused with the ''[[HPS stain]]''.


===Interpretation===
===Interpretation===
Line 327: Line 584:
*Yellow (saffron) = collagen.
*Yellow (saffron) = collagen.


==Jones' stain==
==Jones stain==
*AKA ''PAS methenamine technique''.<ref name=pmid>{{Cite journal  | last1 = Jones | first1 = DB. | title = Nephrotic glomerulonephritis. | journal = Am J Pathol | volume = 33 | issue = 2 | pages = 313-29 | month =  | year =  | doi =  | PMID = 13402889 | PMC = 1934622 }}</ref>
*[[AKA]] ''Methenamine PAS'', abbreviated ''MPAS''.
 
===Use===
===Use===
*Visualize basement membrane in kidney biopsies.
*Visualize basement membrane in kidney biopsies.
**Especially useful for the diagnosis of [[membranous nephropathy]] (MN).


===Interpretation===
===Interpretation===
Line 336: Line 597:
*Pink = other structures/background.
*Pink = other structures/background.


Notes:<ref>[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF]</ref>
Notes:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF]. Accessed on: 19 May 2011.</ref>


==Hale's colloidal iron==
====Images====
===Use===
<gallery>
*[[Renal cell carcinoma]] vs. oncocytoma; +ve in chromophobe [[renal cell carcinoma]].<ref name=pmid9537468>{{cite journal |author=Tickoo SK, Amin MB, Zarbo RJ |title=Colloidal iron staining in renal epithelial neoplasms, including chromophobe renal cell carcinoma: emphasis on technique and patterns of staining |journal=Am. J. Surg. Pathol. |volume=22 |issue=4 |pages=419–24 |year=1998 |month=April |pmid=9537468 |doi= |url=http://meta.wkhealth.com/pt/pt-core/template-journal/lwwgateway/media/landingpage.htm?issn=0147-5185&volume=22&issue=4&spage=419}}</ref>
Image:Membranous_nephropathy_-_mpas_-_very_high_mag.jpg | [[Membranous nephropathy|MN]] demonstrated with a MPAS - very high mag. (WC/Nephron)
Image:Membranous_nephropathy_-_cropped_-_mpas_-_very_high_mag.jpg | MN demonstrated with a MPAS - very high mag. (WC/Nephron)
</gallery>
 
==Hale's colloidal iron stain==


===Interpretation===
{{Main|Hale's colloidal iron stain}}
*Blue is positive.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>


Notes:
==von Kossa stain==
*A few staff think this is a totally useless stain.<ref>ALS several occasions - 2009.</ref>
===General===
*Type of silver stain.<ref name=pmid8360080>{{Cite journal  | last1 = Rungby | first1 = J. | last2 = Kassem | first2 = M. | last3 = Eriksen | first3 = EF. | last4 = Danscher | first4 = G. | title = The von Kossa reaction for calcium deposits: silver lactate staining increases sensitivity and reduces background. | journal = Histochem J | volume = 25 | issue = 6 | pages = 446-51 | month = Jun | year = 1993 | doi =  | PMID = 8360080 }}</ref>


==von Kossa==
===Use===
===Use===
*Look for calcium.
*Look for calcium.
*Actually stains phosphates and carbonates as a surrogate for calcium.


===Interpretation===
===Interpretation===
*Black = calcium.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>
*Black = calcium.<ref name=Ref_WMSP682>{{Ref WMSP|682}}</ref>


==Toluidine blue==
==Toluidine blue stain==
===Use===
===Use===
*May be useful in kidney biopsies.<ref name=pmid16799480>{{cite journal |author=Fischer EG, Moore MJ, Lager DJ |title=Fabry disease: a morphologic study of 11 cases |journal=Mod. Pathol. |volume=19 |issue=10 |pages=1295–301 |year=2006 |month=October |pmid=16799480 |doi=10.1038/modpathol.3800634 |url=http://www.nature.com/modpathol/journal/v19/n10/abs/3800634a.html}}</ref>
*May be useful in kidney biopsies.<ref name=pmid16799480>{{cite journal |author=Fischer EG, Moore MJ, Lager DJ |title=Fabry disease: a morphologic study of 11 cases |journal=Mod. Pathol. |volume=19 |issue=10 |pages=1295–301 |year=2006 |month=October |pmid=16799480 |doi=10.1038/modpathol.3800634 |url=http://www.nature.com/modpathol/journal/v19/n10/abs/3800634a.html}}</ref><ref name=pmid21659728>{{Cite journal  | last1 = Nicholas | first1 = SB. | last2 = Basgen | first2 = JM. | last3 = Sinha | first3 = S. | title = Using stereologic techniques for podocyte counting in the mouse: shifting the paradigm. | journal = Am J Nephrol | volume = 33 Suppl 1 | issue =  | pages = 1-7 | month =  | year = 2011 | doi = 10.1159/000327564 | PMID = 21659728 }}</ref>
*Stains [[mast cell]]s, [[pneumocystis jirovecii]].
*Stains [[mast cell]]s, [[pneumocystis jirovecii]].


===Interpretation===
===Interpretation===
*Dark blue - nuclei, mast cell granules.
*Dark blue - nuclei, mast cell granules (darker than nuclei).
*Light blue - cytoplasm.
*Light blue - cytoplasm.
*Red/magneta - cartilage. (???)


Ref.: looks a bit sketchy - <ref>[http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html]</ref>
Refs: looks a bit sketchy<ref>URL: [http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html]. Accessed on: 17 March 2011.</ref>, <ref>URL: [http://www.dermnetnz.org/doctors/dermatopathology/stains.html http://www.dermnetnz.org/doctors/dermatopathology/stains.html]. Accessed on: 17 March 2011.</ref>


Image:
====Image====
*[http://commons.wikimedia.org/wiki/File:Smear_of_Pneumocystis_carinii._Toluidine_blue_stain_PHIL_596_lores.jpg PCP stained with toluidine blue (mediawiki.org)].
<gallery>
Image:Smear_of_Pneumocystis_carinii._Toluidine_blue_stain_PHIL_596_lores.jpg | [[PCP]] stained with toluidine blue. (WC)
</gallery>
www:
*[http://www.biomedcentral.com/1471-2407/5/121/figure/F3?highres=y Mast cells stained with toluidine blue (biomedcentral.com)].
*[http://www.biomedcentral.com/1471-2407/5/121/figure/F3?highres=y Mast cells stained with toluidine blue (biomedcentral.com)].


Line 374: Line 643:
*Many variants of this stain exist.
*Many variants of this stain exist.
*Specimens are air-dried.
*Specimens are air-dried.


Interpretation:<ref>{{cite journal |author=Horobin RW, Walter KJ |title=Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears |journal=Histochemistry |volume=86 |issue=3 |pages=331–6 |year=1987 |pmid=2437082 |doi= |url=http://www.springerlink.com/content/r81x25451m841866/}}</ref>
Interpretation:<ref>{{cite journal |author=Horobin RW, Walter KJ |title=Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears |journal=Histochemistry |volume=86 |issue=3 |pages=331–6 |year=1987 |pmid=2437082 |doi= |url=http://www.springerlink.com/content/r81x25451m841866/}}</ref>
Line 381: Line 649:
*Purple - nuclear chromatin, neutrophil granules, platelets.
*Purple - nuclear chromatin, neutrophil granules, platelets.


===Field's stain===
===Field stain===
*Variant of the ''Romanowsky stain'' for rapid processing.
*Variant of the ''Romanowsky stain'' for rapid processing.
*Tends to "blow-up" cell, i.e. cells are larger vis-a-vis ''[[Pap stain]]''.
*Tends to "blow-up" cell, i.e. cells are larger vis-a-vis ''[[Pap stain]]''.


===Diff-Quik===
===Diff-Quik===
:Pronounced ''Diff-Quick''.
*Proprietary variant of ''Romanowsky stain''.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm]. Accessed on: 4 January 2010.</ref>
*Proprietary variant of ''Romanowsky stain''.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm]. Accessed on: 4 January 2010.</ref>
*Use: cytopathology.
 
Uses:
*[[Cytopathology]].
*[[Helicobacter gastritis]] - organisms are dark blue against a light blue background.<ref>URL: [http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm]. Accessed on: 30 August 2012.</ref>
 
====Images====
<gallery>
Image: Lung adenocarcinoma - Diff-Quik -- high mag.jpg | [[Pulmonary_cytopathology#Adenocarcinoma|Lung adenocarcinoma]] - DQ - high mag. (WC)
Image: Lung adenocarcinoma - Diff-Quik -- very high mag.jpg | Lung adenocarcinoma - DQ - very high mag. (WC)
Image: Lung small cell carcinoma - Diff-Quik -- very high mag.jpg | Lung SmCC - DQ - very high mag. (WC)
Image: Lung small cell carcinoma - Diff-Quik -- extremely high mag.jpg | Lung SmCC - DQ - extremely high mag. (WC)
</gallery>


===Wright stain===
===Wright stain===
Line 395: Line 675:
*Blood films.
*Blood films.


===May-Grünwald-Giemsa===
===May-Grünwald-Giemsa stain===
*A variant of the ''Romanowsky stain''; popular in Europe.
*A variant of the ''Romanowsky stain''; popular in Europe.
*Abbreviated ''MGG''.
*Abbreviated ''MGG''.
Line 404: Line 684:


==Papanicolaou stain==
==Papanicolaou stain==
*Often abbreviated ''Pap stain''.
*Abbreviated ''Pap stain''.
*Can be thought of as the H&E of cytopathology.
{{Main|Papanicolaou stain}}
**It is a modified H&E stain.
*Specimens are fixed in ethanol.
*Good for seeing nuclear detail.
*Out-of-focus cytoplasm is translucent; allows one to focus overlapped cells in different planes.


===Use===
==Fontana-Masson stain==  
*Cytopathology.
*[[AKA]] ''Masson-Fontana stain'',<ref name=pmid16081962>{{Cite journal  | last1 = Gaitanis | first1 = G. | last2 = Chasapi | first2 = V. | last3 = Velegraki | first3 = A. | title = Novel application of the masson-fontana stain for demonstrating Malassezia species melanin-like pigment production in vitro and in clinical specimens. | journal = J Clin Microbiol | volume = 43 | issue = 8 | pages = 4147-51 | month = Aug | year = 2005 | doi = 10.1128/JCM.43.8.4147-4151.2005 | PMID = 16081962 }}</ref> ''Fontana-Masson stain for melanin'', ''melanin stain''.
{{Main|Fontana-Masson stain}}
 
==Schmorl's stain==
*Stains melanin.
**Similar to ''Fontana-Masson stain''.
 
Notes:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: 5 May 2010.</ref>
 
==Martius scarlet blue stain==
===General===
*Stains connective tissue and fibrin.<ref>URL: [http://www.bris.ac.uk/vetpath/cpl/msb.html http://www.bris.ac.uk/vetpath/cpl/msb.html]. Accessed on: 26 November 2010.</ref>
*Abbreviated ''MSB''.


Use:
*Look for fibrinoid [[necrosis]] in [[vasculitis]].
===Interpretation===
===Interpretation===
*Blue/purple = nucleus.
*Muscle and fibrin - red.
*Green/pink = cytoplasm.
*Nuclei = brown/black.  
*Orange = keratin.
*Collagen - blue.  
*Red blood cells - yellow.  


Image:
Image:
*[http://commons.wikimedia.org/wiki/File:Urine_citology_urothelial_carcinoma_2.jpg Pap stain - urine cytology (wikimedia.org)].
*[https://www.ole.bris.ac.uk/bbcswebdav/institution/Faculty%20of%20Health%20Sciences/Veterinary%20Science/eLearning%20resources/Pathology%20Laboratory%20Protocols/hst/msb.html MSB (bris.ac.uk)].


==Fontana-Masson==
Ref.:<ref>URL: [http://www.bris.ac.uk/vetpath/cpl/msb.html http://www.bris.ac.uk/vetpath/cpl/msb.html]. Accessed on: 26 November 2010.</ref>
*AKA ''Fontana-Masson stain for melanin'', ''melanin stain''.
*A type of silver stain.


Stains:
==Picro-Mallory stain==
#Melanin.
===General===
#"argentaffin granules" of the digestive tract.
*Find fibrin.


===Use===
===Interpretation<ref>{{cite web |url=http://stainsfile.info/StainsFile/stain/fibrin/picro-mallory-1.htm |title=Picro-Mallory for Fibrin – Long Version |author= |date= |work= |publisher= |accessdate=17 January 2011}}</ref>===
*Stain for melanin.
* Fibrin = red.
**Used to differentiate brown pigments (lipofuscin, hemosiderin, melanin).<ref>URL: [http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exfontana.htm http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exfontana.htm]. Accessed on: 5 May 2010.</ref>
* Erythrocytes = yellow.
***Many pathologists prefer IHC, i.e. ''Melan A'' over this stain.
* Connective tissue = blue.


Image:
Image:
*[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exfontana.htm Fontana-Masson stain (vetmed.vt.edu)].
*[http://4.bp.blogspot.com/_rlWIjG4vffU/S3FC7BNIJHI/AAAAAAAACDc/XfOz9X_bvbY/s1600-h/publicado+lengua%233.jpg Picro-Mallary (blogspot.com)].
 
==Verhoeff-van Gieson stain==
:''Verhoeff stain'' redirects here.
*[[AKA]] Elastic van Gieson stain, abbreviated ''EVG''.
===General===
*Similar to ''Masson Trichrome & Verhoeff stain''.<ref>URL: [http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm]. Accessed on: 3 January 2011.</ref>
 
Use:
*Examine large blood vessels.<ref>URL: [http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvvg.htm http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvvg.htm]. Accessed on: 3 January 2011.</ref>
 
===Interpretation===
*Elastin = black.
*Collagen = bright red.
*Muscle = dull red.
 
<gallery>
File:Cerebral aneurysm EVG stain.jpg |EVG stain of a cerebral aneurysm. (WC/jensflorian)
</gallery>
 
==Copper stain==
===General===
*Used in liver biopsies.
*May be seen in [[Wilson's disease]].
 
Note:
*Copper staining is a non-specific finding seen in many liver diseases; it is associated with impaired bile secretion.<ref name=pmid2464523>{{cite journal |author=Miyamura H, Nakanuma Y, Kono N |title=Survey of copper granules in liver biopsy specimens from various liver abnormalities other than Wilson's disease and biliary diseases |journal=Gastroenterol. Jpn. |volume=23 |issue=6 |pages=633–8 |year=1988 |month=December |pmid=2464523 |doi= |url=}}</ref>
 
===Interpretation===
*Copper = red granules.
 
Images:
*[http://www.naika.or.jp/im2/42/10/figs/14/fig1.jpg Wilson's disease (naika.or.jp)].<ref>URL: [http://www.naika.or.jp/im2/42/10/14c.aspx http://www.naika.or.jp/im2/42/10/14c.aspx]. Accessed on: 24 January 2011.</ref>
 
==Shikata stain==
*[[AKA]] Orcein stain for copper-protein.
*[[AKA]] Shikata-Cu,<ref>[http://www.mayomedicallaboratories.com/test-catalog/Overview/9836 http://www.mayomedicallaboratories.com/test-catalog/Overview/9836]. Accessed on: 24 January 2011.</ref>
*[[AKA]] Shikata's orcein staining.<ref>URL: [http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat]. Accessed on: 24 January 2011.</ref>
 
===General===
*Used in [[medical liver disease|medical liver]] biopsies - stains sulfhydrl groups and identifies:
**Copper-associated protein.
**Elastin.
**[[Hepatitis B]] surface antigen.<ref name=pmid7822848>{{Cite journal  | last1 = Ghosh | first1 = AK. | last2 = Dasgupta | first2 = A. | last3 = Raha | first3 = K. | last4 = Jana | first4 = A. | last5 = Majumdar | first5 = DN. | title = Hepatic histology in chronic liver disease in hepatitis B surface antigen positive cases. | journal = J Indian Med Assoc | volume = 92 | issue = 10 | pages = 333-5 | month = Oct | year = 1994 | doi =  | PMID = 7822848 }}</ref>
 
===Interpretation===
Features:<ref>URL: [http://www.nottingham.ac.uk/pathology/protocols/shikata.html http://www.nottingham.ac.uk/pathology/protocols/shikata.html]. Accessed on: 24 January 2011.</ref>
*Dark purple/brown = elastin fibres, HBsAg and copper-associated protein
*Light purple = background
*Red = nuclei (only if counter-stain used)
 
==Gömöri Trichrome stain==
*Named after George Gömöri<ref>GOMORI, G. - A rapid one-step trichrome stain. Am. J. Clin. Path. 20: 661-664, 1950.</ref>


==Schmorl's stain==
===General===
*Stains melanin.
*Used in [[muscle biopsies]] - used to find abnormal mitochondrial deposits.
**Similar to ''Fontana-Masson stain''.
 
===Interpretation===
*Dark green = muscle fibers.
*Red = nuclei.
*Bright red = mitochondria, red blood cells.
 
Images:
<gallery>
File:Ragged red fibers in MELAS.jpg | Ragged red fibers in MELAS, a mitochondrial disease. (WC)
File:Dilated peri-tubular capillaries filled with sickled RBCs, original Gomori's trichrome stain.jpg | Sickle cell nephropathy. (WC)
</gallery>
 
==Miller stain==
===General===
*Stains elastin.
*Used to identify blood vessels and [[pleural invasion]] in [[lung cancer]].
 
===Interpretation===
Staining:<ref>URL: [https://www.ihcworld.com/_protocols/special_stains/miller's_elastic_ellis.htm https://www.ihcworld.com/_protocols/special_stains/miller's_elastic_ellis.htm]. Accessed on: 28 August 2015.</ref>
*Black = elastin fibres, granules in mast cells.
*Red = collagen.
*Yellow = muscle, fibrin, [[erythrocytes]].
*Green/brown = nuclei.


Notes:<ref>URL: [http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html]. Accessed on: 5 May 2010.</ref>
====Images====
<gallery>
Image: Normal visceral pleura of lung - Miller -- high mag.jpg | Miller stain showing lung tissue. (WC)
</gallery>


==See also==
=See also=
*[[Immunohistochemistry]].
*[[Immunohistochemistry]].
*[[Basics]].
*[[Basics]].


==References==
=References=
{{reflist|2}}
{{reflist|2}}


==External links==
=External links=
*[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MANUALS.html Procedure manuals] - med.utah.edu.
*[http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MANUALS.html Procedure manuals] - med.utah.edu.
*[http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html Special stains (introduction)] - med.utah.edu.
*[http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html Special stains (introduction)] - med.utah.edu.
*[http://www.histology-world.com/stains/stains.htm Stains] - histology-world.com.
*[http://www.histology-world.com/stains/stains.htm Stains] - histology-world.com.


[[Category:Basics of pathology]]
[[Category:Basics]]
[[Category:Stains|Stains]]

Latest revision as of 16:05, 1 May 2024

Hematoxylin and eosin stain of benign prostate gland.

This article deals with stains. The H&E stain is only one of many stains that are used in pathology.

Non-H&E stains are often referred to as special stains.

Where to start...

Principles

When considering additional (i.e. special) stains one should (in order) do the following:[1]

  1. Make sure one has exhausted the clinical history; history is considered the best special stain.
  2. Special stains (below).
  3. Immunohistochemistry (dealt with in a separate article).
  4. Molecular testing, electron microscopy.

Common stains

  1. H&E stain.
  2. PAS stain.
  3. PAS-D stain.
  4. AFB stains, e.g. Ziehl-Neelsen stain.
  5. Congo red.
  6. GMS stain.
  7. Gram stain.

Immunohistochemistry

Main article: Immunohistochemistry

General

  • Abbreviated IHC.

Interpretation

Simple version:

  • Positive is (usually): brown.
  • Negative tissue is: light blue.

Important notes:

  • One has to know where the target (of the antibody) is supposed to be, i.e. cytoplasm vs. cell membrane.
  • The edge of the tissue may have light staining - edge effect.
  • If everything is brown... suspect that it didn't work.
  • In some situations you're blessed with an internal control, e.g. in renal tumours CD10 will stain RCC and the proximal tubule, in GISTs - CD117 the mast cells are positive.

Work-up of infection

It often not possible to be definitive by staining.[2]

Basic panel:

  • Gram stain - for bacteria.
  • GMS stain - fungal stain.
  • PAS (or PAS-D) - fungal stain.

Fungi

Main article: Fungi

Fungi are a type of microorganisms. They are seen by pathologist every once in a while.

Specific stains

What follows is a big list... of stains.

Haematoxylin and eosin stain

  • Abbreviated H&E.
Main article: Hematoxylin and eosin stain

Haematoxylin phyloxin saffron stain

General

  • Abbreviated HPS.
  • An alternative to the H&E stain - some pathol. departments use this as their standard.

Interpretation

  • Haematoxylin = blue -- stains nucleus.
  • Phyloxin = pink -- stains muscle and cytoplasm.
  • Saffron = yellow -- stains collagen.
  • An alternative to H&E stain.
    • Fibrosis is easier to see on HPS than H&E... as one can see the collagen.

Images

  • Perineural invasion - prostate - HPS stain (WC)

  • Meningioma - HPS stain (WC)

  • Endoneurial fibrosis - HPS stain (WC)

Periodic acid Schiff stain

  • Abbreviated PAS.

Primary application

  • Kidney biopsies, medical.
  • Liver biopsies, medical.
    • Positive in alpha-1 antitrypsin deficiency.

Utility

  • Stains - lipofuscin,[3] basement membranes, fungi, glycogen, (neutral) mucin.

Interpretation

  • Magenta = glycogen, mucin, fungi.
  • Blue = nuclei.

Ref.:[4]

Image

  • Metanephric adenoma - PAS (WC)

  • Liver biopsy with glycogen storage disease - PAS (WC)

  • Esophageal candidiasis - PAS (WC)

Periodic acid Schiff fungal stain

  • Abbreviated PASF.

Primary application

Interpretation

  • Light purple = fungi.
  • Light green = background.
  • Washed-out light purple = Gram positive bacilli.

Note:

  • This is much improved over the PAS in the context of skin, as the background is similar to the fungal organisms.

  • Fungal organisms. PASF stain.

Periodic acid Schiff with diastase

  • Abbreviated: PAS-D and PASD.

General

  • Diastase digests the glycogen.
  • "PAS diastase resistant"[5] implies PASD +ve and PAS +ve.

Use

  • Stains mucin.
  • Used to identify glycogen (together with PAS stain).
    • Glycogen = clear (digested) on PAS-D.
    • Glycogen = magenta on PAS.

Notes: [6]

Interpretation

  • Light purple = fungi.
  • Light blue/pink = background. ???

Gomori methenamine-silver stain

  • Abbreviated GMS.

Note:

  • GMS is "Grocott's methenamine Silver" according to WMSP.[7]

Use

Image

  • GMS showing histoplasma (WC/Nephron)

Acid-fast bacilli stains

  • Abbreviated: AFB.

There are several AFB stains:

Ziehl-Neelsen stain

  • Most popular acid-fast bacilli stain.
  • Stains other mycobacteria -- not specific for tuberculosis.
    • Stains Nocardia.[10]

Image

  • ZN stain. (WC/CDC)

Fite stain

Interpretation:

  • Red = AFB.
  • Blue = background.

Auramine-rhodamine stain

  • Fluorescent stain.

Image

  • AR stain. (WC/CDC)

Kinyoun stain

Congo red stain

Use

  • Used to look for amyloid.
    • Mnemonic: CRAP = congo red amyloid protein.
  • An alternate stain for amyloid is Thioflavin T.

Note:

  • Thick sections (~10 micrometers) are considered a requirement for the stain to work properly.[13]
    • If the section is too thin... it doesn't work.

Interpretation

  • Amyloid = pink/red.
  • Nuclei = blue.

Ref.:[14]

Image

Thioflavin T stain

Use

Interpretation

  • Amyloid = green.

Image: Amyloid (inano.au.dk).

Gram stain

Use

  • "It is useless for finding bacteria."[15]
    • If they are to be seen... they'll be visible on H&E.

Note:

  • Microbiology is better at finding organisms than pathology.
    • They have one significant advantage -- if a small amount of bugs are present... they grows into a large (obviously visible) colony.

DDx for common patterns

A short list of bacteria and their characteristics:[16]

Shape\Gram stain Positive Negative Variable or negative
Bacilli Clostridium difficile, Bacillus anthracis, Nocardia spp. Escherichia coli, Helicobacter pylori, Yersinia pestis, Hemophilus influenzae Mycobacterium tuberulosis, Legionella pneumophila[17]
Cocci Streptococcus pneumoniae, Staphylococcus aureus Neisseria meningitidis, Moraxella catarrhalis

Interpretation

  • Purple (or blue) = Gram positive organisms.
  • Red = Gram negative organisms, nuclei.[18]
  • Yellow = background.

Notes:

  • Many of the bacteria are quite small relative to lymphocytes; Escherichia coli is 1-2 micrometers long x 0.25 micrometers in diameter.[19]
  • Epithelial cell nuclei & stromal cell nuclei may stain red.
  • Memory device: purple = positive.

Images

  • Gram positive cocci. (WC)

  • Gram positive rods - anthrax. (WC)

Luxol fast blue stain

  • Abbreviated LFB.

Use

  • Neuropathology, myelin stain.

Intepretation

  • Blue = myelinated fibers (contain lipoproteins), lipofuscin.[20]
    • Lack of blue (where it ought to be) = demyelination.
  • Purple = nerve cell (e.g. neuron).
  • Neutrophils = pink.

Ref.:[21]

Image

  • Globus pallidus and putamen - H&E-LFB. (WC)

  • Normal cortex - LFB only. (WC/jensflorian)

  • White-grey matter junction - LFB. (WC/jensflorian)

Giemsa stain

Use

Interpretation

  • Tissue is light blue/green.
  • Goblet cells are purple.[23]

Image:

Reticulin stain

Use

  • Liver biopsy, medical.
    • Demonstrates the reticular fibers (in cirrhosis the fibers are disrupted).
  • Before IHC, reticulin was used to differentiate sarcomas from carcinomas:[25]
    • Sarcomas have reticulin around each cell.
    • Carcinomas have reticulin around clusters of cells.
  • Commonly used in neuropathology.
    • In adenoma, reticulin highlights the lost acinar structure of normal pituitary gland.
    • Paraganglioma (Zellballen architecture)
    • Separating schwannoma (basement membrane around each cell) from meingioma in cerebellopontine angle.
    • Separating desmoplastic medulloblastoma from classic/anaplastic forms.


Interpretation

  • Black = reticular fibers.
  • Red = nuclei.

Notes:[26]

Images

  • Liver. Reticulin stain. (WC)

  • Hepatic adenoma. Reticulin stain. (WC)

  • Reticulin stain highlighting the "Zellballen" architecture of paraganglioma. (WC/jensflorian)

  • Reticulin staina round the "pale islands" of a desmoplastic medulloblastoma. (WC/jensflorian)

Cresyl violet stain

Use

  • Used at some places (e.g. SMH) to look for Helicobacter organisms.

Interpretation

  • Everything is shades of blue.
    • Helicobacter stains blue.

Prussian blue stain

  • AKA Perl's iron stain.

Use

  • Useful for iron and hemosiderin; useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).

Interpretation

  • Blue = iron.

Image:

Notes:

  • Described well by vetmed.vt.edu.[27]
  • DDx of brown pigment: Fontana-Masson (melanin), Kluver-Barrera stain (lipofuscin).

Images

  • Liver hemosiderosis. Prussian blue stain. (WC/Nephron)

  • CSF Siderophages in subarachnoid hemorrhage. (WC/jensflorian)

Kluver-Barrera stain

Combination of:

  • Luxol Fast Blue,
  • Cresyl Violet,
  • Special component for lipofuscin.

Use

  • Useful for differentiating brown pigments (melanin, lipofuscin, tattoo pigment, hemosiderin).
    • Stains lipofuscin.
  • Useful to detect demyelinating lesions in the CNS.

Notes:

  • PAS also stains lipofuscin and is more commonly available.

Interpretation

  • Blue pigmented granules = lipofuscin.

Notes:

  • Described well by vetmed.vt.edu.[28]
  • DDx of brown pigment: Fontana-Masson (melanin), Prussian blue stain (hemosiderin).

  • Encephalomyelitis disseminata (Klüver-Barrera)

Oil red O stain

Use

  • Stains adipose tissue.
  • Corroborate diagnosis of lipoid pneumonia.[29]
  • Screen for GERD - positive staining seen in macrophages from BAL specimens.[30]
  • Uncommon.

Notes:

  • Must be done on fresh tissue, i.e. it cannot be fixed in formalin.

Interpretation

  • Red = fat.

Images

  • Oil red O stain. (WC)

Warthin-Starry stain

Background:

  • Developed by a bunch of pathologists in Michigan to look for spirochetes.[31]

Use

  • Find spirochetes, e.g. syphilis (Treponema pallidum),[32] cat-scratch disease (Bartonella henselae).
  • Find Helicobacter spp., e.g. Helicobacter pylori -- Mount Sinai Hospital.[33]

Interpretation:[34]

  • Spirochetes - black.
  • Background - yellow.

Image

  • Helicobacter gastritis - Warthin-Starry stain. (WC)

Notes:

  • Considered a "dirty" stain - picks-up junk in the background.[35]

Dieterle stain

Considered a variant of the Steiner stain.[36]

Use

  • Find spirochetes, e.g. syphilis (Treponema pallidum),[37] donovan bodies (leishmaniasis),[38] Helicobacter pylori and Bartonella henselae (Cat-scratch disease).[39]

Interpretation

  • Spirochetes - black.
  • Background - yellow.

Images

  • Dieterle stain - T. pallidum. (WC)

www:

Bielschowsky stain

Abbreviated: Biel stain.

Use

  • Stains glial tissue, i.e. brain.
    • Demonstrates neurofibrillary tangles, senile plaques (as in Alzheimer's disease).

Interpretation

  • Black = axons, tangles, plaques.
  • Brown/dark brown = plaque, vascular amyloid.
  • Yellow/brown = other.

Ref.: [40]

Image

  • Bielschowsky stain. (WC/Nephron)

Mucicarmine stain

  • Stains some mucins... uses the dye carmine.

Use

  • Identify mucin.
  • Malignant cells that produce mucin... carcinomas.[41]

Interpretation

  • Carmine with metanil yellow and Weigert's Hematoxylin:[42]
    • Blue/black = nucleus.
    • Yellow = background.
    • Red = mucin.[43]

Images

www:

Alcian blue stain

General

  • Stains acidic mucin (pH=2.5); Alcian blue = Acidic.
    • A variant uses pH=1.0.[7]

Note:

  • Alcian blue (not otherwise specified) usu. refers to the pH=2.5.[44]

Use

Note:

  • Esophageal submucosal glands - alcian blue positive.

Interpretation

  • Blue = acidic mucins.[45]

Notes:

Image

Sodium sulphate-alcian blue stain

Sulfated alcian blue (abbreviated 'SAB) redirects here.

Use

Interpretation

  • Green = amyloid.[46]
    • Other things that are green: mast cells, mucoid degeneration, basophilic myofibre degeneration, califications.
  • Yellow = background.

Image:

Movat's stain

Use

Components

  • Acid fuchsin, alcian blue, crocein scarlet, elastic hematoxylin, and saffron.[48][49]

Interpretation of Movat stain

  • Black = nuclei and elastic fibers.
  • Yellow = collagen and reticular fibers.
  • Blue = mucin, ground substance.
  • Red (intense) = fibrin.
  • Red = muscle.

Reference: [49]

How to remember? A.: Primary colours (red, blue, yellow) + black.

Images

  • Cardiac amyloidosis - Movat stain. (WC/Nephron)

  • Cystic medial degeneration - Movat stain - low mag. (WC/Nephron)

  • Cystic medial degeneration - Movat stain - intermed. mag. (WC/Nephron)

  • Cystic medial degeneration - Movat stain - high mag. (WC/Nephron)

Masson's trichrome stain

General

  • Collagen vs. muscle.

Interpretation

  • Black = nuclei.
  • Red = muscle (smooth muscle actin).
  • Baby blue = collagen.
  • Blue gray = amyloid.[50]

Notes: [51]

Elastic trichrome stain

General:

  • "Elastic trichrome" is one important variant of Masson's trichrome.

Interpretation - as above in Masson's trichrome - plus:

  • Black = nuclei and elastin.

Mallory trichome stain

General

  • Collagen vs. muscle.
  • May be done with elastin.

Site

  • Kidney Bx (to assess for fibrosis).
  • Liver Bx (to assess for cirrhosis).
  • Cardiovascular/lung (to see differentiate the layers of the arteries, and arteries from veins).

Interpretation

  • Black = nuclei.
  • Red = muscle (smooth muscle actin).
  • Green = collagen.

Image

Haematoxylin orcein phyloxin saffron stain

  • Abbreviated HOPS.[52]
  • It should not be confused with the HPS stain.

Interpretation

  • Blue (haematoxylin) = nuclei.
  • Black (orcein) = elastin.
  • Red (phyloxin) = muscle.
  • Yellow (saffron) = collagen.

Jones stain

  • AKA PAS methenamine technique.[53]
  • AKA Methenamine PAS, abbreviated MPAS.

Use

  • Visualize basement membrane in kidney biopsies.

Interpretation

  • Black = basement membrane.
  • Blue = nuclei.
  • Pink = other structures/background.

Notes:[54]

Images

  • MN demonstrated with a MPAS - very high mag. (WC/Nephron)

  • MN demonstrated with a MPAS - very high mag. (WC/Nephron)

Hale's colloidal iron stain

Main article: Hale's colloidal iron stain

von Kossa stain

General

  • Type of silver stain.[55]

Use

  • Look for calcium.
  • Actually stains phosphates and carbonates as a surrogate for calcium.

Interpretation

  • Black = calcium.[7]

Toluidine blue stain

Use

Interpretation

  • Dark blue - nuclei, mast cell granules (darker than nuclei).
  • Light blue - cytoplasm.
  • Red/magneta - cartilage. (???)

Refs: looks a bit sketchy[58], [59]

Image

  • PCP stained with toluidine blue. (WC)

www:

Romanowsky stain

  • Occasionally spelled Romanowski.
  • Many variants of this stain exist.
  • Specimens are air-dried.

Interpretation:[60]

  • Red - RBCs, eosinophil granules.
  • Blue (basophilic) - lymphocyte cytoplasm.
  • Purple - nuclear chromatin, neutrophil granules, platelets.

Field stain

  • Variant of the Romanowsky stain for rapid processing.
  • Tends to "blow-up" cell, i.e. cells are larger vis-a-vis Pap stain.

Diff-Quik

Pronounced Diff-Quick.
  • Proprietary variant of Romanowsky stain.[61]

Uses:

Images

  • Lung adenocarcinoma - DQ - high mag. (WC)

  • Lung adenocarcinoma - DQ - very high mag. (WC)

  • Lung SmCC - DQ - very high mag. (WC)

  • Lung SmCC - DQ - extremely high mag. (WC)

Wright stain

  • A variant of the Romanowsky stain; popular in North American.

Use:

  • Blood films.

May-Grünwald-Giemsa stain

  • A variant of the Romanowsky stain; popular in Europe.
  • Abbreviated MGG.

Use:

  • Blood films.
  • Cytopathology.

Papanicolaou stain

  • Abbreviated Pap stain.
Main article: Papanicolaou stain

Fontana-Masson stain

  • AKA Masson-Fontana stain,[63] Fontana-Masson stain for melanin, melanin stain.
Main article: Fontana-Masson stain

Schmorl's stain

  • Stains melanin.
    • Similar to Fontana-Masson stain.

Notes:[64]

Martius scarlet blue stain

General

  • Stains connective tissue and fibrin.[65]
  • Abbreviated MSB.

Use:

Interpretation

  • Muscle and fibrin - red.
  • Nuclei = brown/black.
  • Collagen - blue.
  • Red blood cells - yellow.

Image:

Ref.:[66]

Picro-Mallory stain

General

  • Find fibrin.

Interpretation[67]

  • Fibrin = red.
  • Erythrocytes = yellow.
  • Connective tissue = blue.

Image:

Verhoeff-van Gieson stain

Verhoeff stain redirects here.
  • AKA Elastic van Gieson stain, abbreviated EVG.

General

  • Similar to Masson Trichrome & Verhoeff stain.[68]

Use:

  • Examine large blood vessels.[69]

Interpretation

  • Elastin = black.
  • Collagen = bright red.
  • Muscle = dull red.

  • EVG stain of a cerebral aneurysm. (WC/jensflorian)

Copper stain

General

Note:

  • Copper staining is a non-specific finding seen in many liver diseases; it is associated with impaired bile secretion.[70]

Interpretation

  • Copper = red granules.

Images:

Shikata stain

  • AKA Orcein stain for copper-protein.
  • AKA Shikata-Cu,[72]
  • AKA Shikata's orcein staining.[73]

General

  • Used in medical liver biopsies - stains sulfhydrl groups and identifies:

Interpretation

Features:[75]

  • Dark purple/brown = elastin fibres, HBsAg and copper-associated protein
  • Light purple = background
  • Red = nuclei (only if counter-stain used)

Gömöri Trichrome stain

  • Named after George Gömöri[76]

General

Interpretation

  • Dark green = muscle fibers.
  • Red = nuclei.
  • Bright red = mitochondria, red blood cells.

Images:

  • Ragged red fibers in MELAS, a mitochondrial disease. (WC)

  • Sickle cell nephropathy. (WC)

Miller stain

General

Interpretation

Staining:[77]

  • Black = elastin fibres, granules in mast cells.
  • Red = collagen.
  • Yellow = muscle, fibrin, erythrocytes.
  • Green/brown = nuclei.

Images

  • Miller stain showing lung tissue. (WC)

See also

References

  1. ↑ LAE. 13 July 2010.
  2. ↑ Woods GL, Walker DH (July 1996). "Detection of infection or infectious agents by use of cytologic and histologic stains". Clin. Microbiol. Rev. 9 (3): 382-404. PMC 172900. PMID 8809467. http://cmr.asm.org/cgi/pmidlookup?view=long&pmid=8809467.
  3. ↑ Kovi J, Leifer C (July 1970). "Lipofuscin pigment accumulation in spontaneous mammary carcinoma of A/Jax mouse". J Natl Med Assoc 62 (4): 287–90. PMC 2611776. PMID 5463681. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2611776/pdf/jnma00512-0077.pdf.
  4. ↑ http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PAS.PDF
  5. ↑ Qizilbash, A.; Young-Pong, O. (Jun 1983). "Alpha 1 antitrypsin liver disease differential diagnosis of PAS-positive, diastase-resistant globules in liver cells.". Am J Clin Pathol 79 (6): 697-702. PMID 6189389.
  6. ↑ http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/PASD.PDF
  7. ↑ 7.0 7.1 7.2 Humphrey, Peter A; Dehner, Louis P; Pfeifer, John D (2008). The Washington Manual of Surgical Pathology (1st ed.). Lippincott Williams & Wilkins. pp. 682. ISBN 978-0781765275.
  8. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/FITES.PDF. Accessed on: 19 May 2011.
  9. ↑ Ilyas S, Youssef D, Chaudhary H, Al-Abbadi MA (September 2011). "Myocbacterium-avium intracellulare associated inflammatory pseudotumor of the anterior nasal cavity". Head Neck Pathol 5 (3): 296–301. doi:10.1007/s12105-011-0248-0. PMC 3173547. PMID 21327589. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3173547/.
  10. ↑ URL: http://library.med.utah.edu/WebPath/LUNGHTML/LUNG024.html. Accessed on: 19 May 2011.
  11. ↑ Kehl, KS.; Cicirello, H.; Havens, PL. (Feb 1995). "Comparison of four different methods for detection of Cryptosporidium species.". J Clin Microbiol 33 (2): 416-8. PMID 7536216.
  12. ↑ Ignatius, R.; Lehmann, M.; Miksits, K.; Regnath, T.; Arvand, M.; Engelmann, E.; Futh, U.; Hahn, H. et al. (Feb 1997). "A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens.". J Clin Microbiol 35 (2): 446-9. PMID 9003613.
  13. ↑ URL: http://www.ihcworld.com/_protocols/special_stains/congo_red_bennhold.htm. Accessed on: 26 January 2012.
  14. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/CONGORED.PDF. Accessed on: 4 December 2010.
  15. ↑ St. Michael's Hospital - Stains Handout.
  16. ↑ URL: http://www.atsu.edu/faculty/chamberlain/Website/pnebact.htm. Accessed on: 7 May 2013.
  17. ↑ URL: http://meded.ucsd.edu/isp/1999/CAP/legion.html. Accessed on: 7 May 2013.
  18. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/GRAM.PDF. Accessed on: 7 May 2013.
  19. ↑ URL: http://www.lpi.usra.edu/publications/slidesets/marslife/slide_27.html.
  20. ↑ MUN. 26 November 2010.
  21. ↑ http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/LFB.PDF
  22. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html. Accessed on: April 6, 2009.
  23. ↑ URL: http://www.kennedy.ox.ac.uk/facilities/histology/histology-information. Accessed on: 17 August 2015.
  24. ↑ URL: http://amser.org/index.php?P=AMSER--ResourceFrame&resourceId=6018. Accessed on: 17 August 2015.
  25. ↑ MACKENZIE DH (March 1958). "Reticulin patterns in the diagnosis of carcinomas and sarcomas". Br. J. Cancer 12 (1): 14–9. PMC 2074006. PMID 13536209. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2074006/.
  26. ↑ http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/RETIC.PDF
  27. ↑ Prussian blue stain. URL:[http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exprussb.htm. Accessed on: 5 May 2010.
  28. ↑ Kluver-Barrera stain. URL:http://education.vetmed.vt.edu/curriculum/VM8054/labs/Lab2/Examples/exkluvbarr.htm. Accessed on: 5 May 2010.
  29. ↑ Yampara Guarachi, GI.; Barbosa Moreira, V.; Santos Ferreira, A.; Sias, SM.; Rodrigues, CC.; Teixeira, GH. (2014). "Lipoid pneumonia in a gas station attendant.". Case Rep Pulmonol 2014: 358761. doi:10.1155/2014/358761. PMID 25374742.
  30. ↑ Hopkins, PM.; Kermeen, F.; Duhig, E.; Fletcher, L.; Gradwell, J.; Whitfield, L.; Godinez, C.; Musk, M. et al. (Aug 2010). "Oil red O stain of alveolar macrophages is an effective screening test for gastroesophageal reflux disease in lung transplant recipients.". J Heart Lung Transplant 29 (8): 859-64. doi:10.1016/j.healun.2010.03.015. PMID 20466562.
  31. ↑ URL: http://www.merriam-webster.com/medical/warthin. Accessed on: 17 August 2010.
  32. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html. Accessed on: April 6, 2009.
  33. ↑ http://www.dako.co.uk/index/prod_search/prod_products.htm?productareaid=41&baseprodidver=A224462007
  34. ↑ http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAIN029.html
  35. ↑ DB. 4 August 2010.
  36. ↑ URL: http://www.mayomedicallaboratories.com/test-catalog/Overview/80327. Accessed on: 8 August 2010.
  37. ↑ Humphrey, Peter A; Dehner, Louis P; Pfeifer, John D (2008). The Washington Manual of Surgical Pathology (1st ed.). Lippincott Williams & Wilkins. pp. 455. ISBN 978-0781765275.
  38. ↑ URL: http://www.mondofacto.com/facts/dictionary?Dieterle%27s+stain. Accessed on: 4 August 2010.
  39. ↑ URL: http://www.mayomedicallaboratories.com/test-catalog/Overview/80327. Accessed on: 8 August 2010.
  40. ↑ http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BIELSCH.PDF
  41. ↑ Lefkowitch, Jay H. (2006). Anatomic Pathology Board Review (1st ed.). Saunders. pp. 681 (Q25). ISBN 978-1416025887.
  42. ↑ Humphrey, Peter A; Dehner, Louis P; Pfeifer, John D (2008). The Washington Manual of Surgical Pathology (1st ed.). Lippincott Williams & Wilkins. pp. 678. ISBN 978-0781765275.
  43. ↑ http://www.medschool.lsuhsc.edu/pathology/pathist/SURGPATH/special%20stains/assets/mucicarmine3.jpg
  44. ↑ URL: http://www.pathologyoutlines.com/topic/stainsalcianblue.html. Accessed on: 11 October 2012.
  45. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/ALCIAN.PDF. Accessed on: 20 December 2011.
  46. ↑ 46.0 46.1 Pomerance, A.; Slavin, G.; McWatt, J. (Jan 1976). "Experience with the sodium sulphate-Alcian Blue stain for amyloid in cardiac pathology.". J Clin Pathol 29 (1): 22-6. PMID 55419.
  47. ↑ URL: http://www.polyrnd.com/products/reagent-assembly-kits/conventional/amyloid-stain---sulfated-alcian-blue-(sab).aspx. Accessed on: October 15, 2014.
  48. ↑ [1]
  49. ↑ 49.0 49.1 Modified Movat's Pentachrome Stain. University Penn Medicine. URL: http://www.med.upenn.edu/mcrc/histology_core/movat.shtml. Accessed on: January 29, 2009.
  50. ↑ Kunnath-Velayudhan S, Larsen BT, Coley SM, De Michele S, Santoriello D, Colby TV, Bhagat G, Saqi A (March 2021). "Masson Trichrome and Sulfated Alcian Blue Stains Distinguish Light Chain Deposition Disease From Amyloidosis in the Lung". Am J Surg Pathol 45 (3): 405–413. doi:10.1097/PAS.0000000000001593. PMID 33002919.
  51. ↑ ULR: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MASSONS.PDF. Accessed on: 2 November 2011.
  52. ↑ Perry JR, Bilbao JM, Gray T (1992). "Fatal basilar vasculopathy complicating bacterial meningitis". Stroke 23 (8): 1175–8. PMID 1636194. Free Full Text.
  53. ↑ Jones, DB.. "Nephrotic glomerulonephritis.". Am J Pathol 33 (2): 313-29. PMC 1934622. PMID 13402889. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1934622/.
  54. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/JONES.PDF. Accessed on: 19 May 2011.
  55. ↑ Rungby, J.; Kassem, M.; Eriksen, EF.; Danscher, G. (Jun 1993). "The von Kossa reaction for calcium deposits: silver lactate staining increases sensitivity and reduces background.". Histochem J 25 (6): 446-51. PMID 8360080.
  56. ↑ Fischer EG, Moore MJ, Lager DJ (October 2006). "Fabry disease: a morphologic study of 11 cases". Mod. Pathol. 19 (10): 1295–301. doi:10.1038/modpathol.3800634. PMID 16799480. http://www.nature.com/modpathol/journal/v19/n10/abs/3800634a.html.
  57. ↑ Nicholas, SB.; Basgen, JM.; Sinha, S. (2011). "Using stereologic techniques for podocyte counting in the mouse: shifting the paradigm.". Am J Nephrol 33 Suppl 1: 1-7. doi:10.1159/000327564. PMID 21659728.
  58. ↑ URL: http://www.molecularstation.com/protocol-links/articles/Toluidine-Blue-Stain-32.html. Accessed on: 17 March 2011.
  59. ↑ URL: http://www.dermnetnz.org/doctors/dermatopathology/stains.html. Accessed on: 17 March 2011.
  60. ↑ Horobin RW, Walter KJ (1987). "Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears". Histochemistry 86 (3): 331–6. PMID 2437082. http://www.springerlink.com/content/r81x25451m841866/.
  61. ↑ URL: http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm. Accessed on: 4 January 2010.
  62. ↑ URL: http://www.ihcworld.com/_protocols/special_stains/diff_quick_ellis.htm. Accessed on: 30 August 2012.
  63. ↑ Gaitanis, G.; Chasapi, V.; Velegraki, A. (Aug 2005). "Novel application of the masson-fontana stain for demonstrating Malassezia species melanin-like pigment production in vitro and in clinical specimens.". J Clin Microbiol 43 (8): 4147-51. doi:10.1128/JCM.43.8.4147-4151.2005. PMID 16081962.
  64. ↑ URL: http://library.med.utah.edu/WebPath/HISTHTML/STAINS/STAINS.html. Accessed on: 5 May 2010.
  65. ↑ URL: http://www.bris.ac.uk/vetpath/cpl/msb.html. Accessed on: 26 November 2010.
  66. ↑ URL: http://www.bris.ac.uk/vetpath/cpl/msb.html. Accessed on: 26 November 2010.
  67. ↑ "Picro-Mallory for Fibrin – Long Version". http://stainsfile.info/StainsFile/stain/fibrin/picro-mallory-1.htm. Retrieved 17 January 2011.
  68. ↑ URL: http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvrmass.htm. Accessed on: 3 January 2011.
  69. ↑ URL: http://education.vetmed.vt.edu/Curriculum/VM8054/Labs/Lab2/Examples/exvvg.htm. Accessed on: 3 January 2011.
  70. ↑ Miyamura H, Nakanuma Y, Kono N (December 1988). "Survey of copper granules in liver biopsy specimens from various liver abnormalities other than Wilson's disease and biliary diseases". Gastroenterol. Jpn. 23 (6): 633–8. PMID 2464523.
  71. ↑ URL: http://www.naika.or.jp/im2/42/10/14c.aspx. Accessed on: 24 January 2011.
  72. ↑ http://www.mayomedicallaboratories.com/test-catalog/Overview/9836. Accessed on: 24 January 2011.
  73. ↑ URL: http://informahealthcare.com/doi/abs/10.3109/00313027709085239?journalCode=pat. Accessed on: 24 January 2011.
  74. ↑ Ghosh, AK.; Dasgupta, A.; Raha, K.; Jana, A.; Majumdar, DN. (Oct 1994). "Hepatic histology in chronic liver disease in hepatitis B surface antigen positive cases.". J Indian Med Assoc 92 (10): 333-5. PMID 7822848.
  75. ↑ URL: http://www.nottingham.ac.uk/pathology/protocols/shikata.html. Accessed on: 24 January 2011.
  76. ↑ GOMORI, G. - A rapid one-step trichrome stain. Am. J. Clin. Path. 20: 661-664, 1950.
  77. ↑ URL: https://www.ihcworld.com/_protocols/special_stains/miller's_elastic_ellis.htm. Accessed on: 28 August 2015.

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